The beneficial use of lipolytic enzymes active on glycolipids in bread making was taught in EP 1 193 314. It was taught that the partial hydrolysis products the lyso-glycolipids were found to have very high emulsifier functionality. However, the enzymes taught in EP 1 193 314 were also found to have significant non-selective activity on triglycerides which resulted in unnecessarily high free fatty acid.
A lipolytic enzyme from Fusarium oxysporum having phospholipase activity has been taught in EP 0 869 167. This lipolytic enzyme has high triacylglyceride hydrolysing (lipase) activity. This enzyme is now sold by Novozymes A/S (Denmark) as Lipopan F™.
WO02/00852 discloses five lipase enzymes and their encoding polynucleotides, isolated from Fusarium venenatum, F. sulphureum, Aspergillus berkeleyanum, F. culmorum and F. solani. All five enzymes are described as having triacylglycerol hydrolysing activity, phospholipase and galactolipase activity.
Lipolytic enzyme variants, with specific amino acid substitutions and fusions, have been produced; some of which have an enhanced activity on the polar lipids compared to the wildtype parent enzymes. WO01/39602 describes such a variant, referred to as SP979, which is a fusion of the Thermomyces lanuginosus lipase, and the Fusarium oxysporum lipase described in EP 0 869 167. This variant has been found to have a significantly high ratio of activity on phospholipids and glycolipids compared to triglycerides.
In WO02/094123 it was discovered that by selecting lipolytic enzymes which were active on the polar lipids (glycolipids and phospholipids) in a dough, but substantially not active on triglycerides or 1-mono-glycerides an improved functionality could be achieved.
In co-pending PCT application number PCT/IB2005/000875, wild-type lipolytic enzymes having a higher ratio of activity on polar lipids as compared with triglycerides are taught. However, this document does not teach lipolytic enzymes from Streptomyces, Thermobifida or Corynebacterium species.
Prior to the present invention no lipolytic enzymes having activity or significant activity on glycolipids had been published from Streptomyces species. Likewise, no lipolytic enzymes having activity or significant activity on glycolipids had been published from Thermobifida species or Corynebacterium species. Although lipases, i.e. triacylglycerol hydrolysing enzymes, have been isolated from Streptomyces species (see Vujaklija et al Arch Microbiol (2002) 178: 124-130 for example), these enzymes have never been identified as having glycolipid hydrolysing activity.